1887

Abstract

The flavin adenine dinucleotide (FAD)-containing putrescine oxidase of catalyses the oxidative deamination of putrescine. The amino acid sequences of the NH-termini of the mature enzyme and lysyl-endopeptidase-generated fragments were determined for preparation of synthetic oligonucleotides as hybridization probes for cloning. A 4·4 kb HI fragment which contained DNA sequences hybridizing to the probes was cloned in pUC19 in . The nucleotide sequence together with the determined amino acid sequences revealed that this enzyme consists of 480 amino acids ( 52000) and contains an FAD-binding consensus sequence at its NH-terminal portion. In front of the transcriptional start point, which is 28 bases upstream of the initiation codon as determined by primer extension, −35 and −10 sequences similar to typical prokaryotic promoter consensus sequences are present. JM109 containing the putrescine oxidase gene just downstream of the promoter in pUC18 produced a large amount of this protein when grown at 37 °C but in the enzymically inactive form of inclusion bodies. However, cultivation of the recombinant cells at temperatures below 30°C led to production of active enzyme (20 times as much as produced by the original strain).

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1993-03-01
2024-03-29
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References

  1. Adachi O., Yamada H., Ogata K. 1966; Purification and properties of putrescine oxidase of Micrococcus rubens. Agricultural and Biological Chemistry 30:1202–1210
    [Google Scholar]
  2. Bibb M. J., Findlay P. R., Johnson M. W. 1984; The relationship between base composition and codon usage in bacterial genes and its use for the simple and reliable identification of protein-coding sequences. Gene 30:157–166
    [Google Scholar]
  3. Chen C. W., Thomas C. A. Jr 1980; Recovery of DNA segments from agarose gel. Analytical Biochemistry 101:339–341
    [Google Scholar]
  4. Desa R. J. 1972; Putrescine oxidase from Micrococcus rubens: purification and properties of the enzyme. Journal of Biological Chemistry 247:5527–5534
    [Google Scholar]
  5. Green M. R., Roeder R. G. 1980; Definition of a novel promoter for the major adenovirus-associated virus mRNA. Cell 22:231–242
    [Google Scholar]
  6. Grunstein M., Hogness D. S. 1975; Colony hybridization: a method for the isolation of cloned DNAs that contain a specific gene. Proceedings of the National Academy of Sciences of the United States of America 723961–3965
    [Google Scholar]
  7. Horinouchi S., Furuya K., Nishiyama M., Suzuki H., Beppu T. 1987; Nucleotide sequence of the streptothricin acetyltransferase gene from Streptomyces lavendulae and its expression in heterologous hosts. Journal of Bacteriology 169:1929–1937
    [Google Scholar]
  8. Kawaguchi Y., Yanagida N., Uozumi T., Beppu T. 1986; Improved direct expression of prochymosin cDNA through changing the SD-ATG codon length. Agricultural and Biological Chemistry 50:499–500
    [Google Scholar]
  9. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature; London: 227680–685
    [Google Scholar]
  10. Maniatis T., Fritsch E. F., Sambrook J. 1982 Molecular Cloning: A Laboratory Manual Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.;
    [Google Scholar]
  11. Mann P. J. G. 1961; Further purification and properties of the amine oxidase of pea seedlings. Biochemical Journal 79:623–631
    [Google Scholar]
  12. Massay V., Palmer G., Bennett R. 1961; The purification and properties of d-amino acid oxidase. Biochimica et Biophysica Acta 48:1–9
    [Google Scholar]
  13. Messing J. 1983; New M13 vectors for cloning. Methods in Enzymology 101:20–78
    [Google Scholar]
  14. Miller J. H. 1972 Experiments in Molecular Genetics Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.;
    [Google Scholar]
  15. Mondovi B., Rotilio G., Finazzi A., Scioscia-Santoro A. 1964; Purification of pig kidney diamine oxidase and its identity with histaminase. Biochemical Journal 91:408–415
    [Google Scholar]
  16. Rosenberg M., Court D. 1979; Regulatory sequences involved in promotion and termination of RNA transcription. Annual Review of Genetics 13:319–353
    [Google Scholar]
  17. Russell D. H., Levy C. C., Schimpff S. C., Hawk I. A. 1971; Urinary polyamines in cancer patients. Cancer Research 31:1555–1558
    [Google Scholar]
  18. Saito H., Miura K. 1963; Preparation of transforming deoxyribonucleic acid by phenol treatment. Biochimica et Biophysica Acta 72:619–629
    [Google Scholar]
  19. Sanger F., Nicklen S., Coulson A. R. 1977; DNA sequencing with chain-terminating inhibitors. Proceedings of the National Academy of Sciences of the United States of America 745463–5467
    [Google Scholar]
  20. Schein C. H., Noteborn M. H. M. 1988; Formation of soluble recombinant proteins in Escherichia coli is favored by lower growth temperature. Bio/Technology 6:291–294
    [Google Scholar]
  21. Scrutton N. S., Berry A., Perham R. N. 1990; Redesign of coenzyme specificity of a dehydrogenase by protein engineering. Nature; London: 34338–43
    [Google Scholar]
  22. Shepard H. M., Yelverton E., Goeddel D. V. 1982; Increased synthesis in E. coli of fibloblast and leukocyte interferons through alterations in ribosome binding site. DNA 1:125–131
    [Google Scholar]
  23. Shine J., Dalgarno L. 1974; The 3‘-terminal sequence of E.coli 16S ribosomal RNA: complementarity to nonsense triplets and ribosome binding sites. Proceedings of the National Academy of Sciences of the United States of America 711342–1346
    [Google Scholar]
  24. Southern E. M. 1975; Detection of specific sequences among DNA fragments separated by gel electrophoresis. Journal of Molecular Biology 98:503–517
    [Google Scholar]
  25. Swoboda B. E. P., Massay V. 1965; Purification and properties of the glucose oxidase from Aspergillus niger. Journal of Biological Chemistry 240:2209–2215
    [Google Scholar]
  26. Tabor H. 1951; Diamine oxidase. Journal of Biological Chemistry 188:125–136
    [Google Scholar]
  27. Takami H., Romsdahl M. M., Nishioka K. 1979; Polyamines in blood-cells as a cancer marker. Lancet ii:912
    [Google Scholar]
  28. Wellner D., Meister A. 1961; Studies on the mechanism of action of l-amino acid oxidase. Journal of Biological Chemistry 236:2357–2364
    [Google Scholar]
  29. Wierenga R. K., De Maeyer M. C. H., Hol W. G. J. 1985; Interaction of pyrophosphate moieties with α-helixes in dinucleotide binding proteins. Biochemistry 24:1346–1357
    [Google Scholar]
  30. Yamada H., Adachi O., Ogata K. 1965a; Putrescine oxidase, a diamine oxidase requiring flavin adenine dinucleotide. Agricultural and Biological Chemistry 29:1148–1149
    [Google Scholar]
  31. Yamada H., Tanaka A., Ogata K. 1965b; Putrescine oxidase of Micrococcus rubens. Agricultural and Biological Chemistry 29:260–261
    [Google Scholar]
  32. Yanisch-Perron C., Vieira J., Messing J. 1985; Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M1M13mp18 and pUC19 vectors. Gene 33:103–109
    [Google Scholar]
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